单克隆抗体检测胃癌前及恶性病变中SAPCD2表达的制备及应用

IF 1.1 4区 医学 Q4 IMMUNOLOGY Iranian Journal of Immunology Pub Date : 2023-05-31 DOI:10.22034/iji.2023.97498.2519
Fang Gao, Yuan Chen, Wenwei Zhang, Xiaolong Wang, Jingchun Li, Feng Feng, Chunli Liu, Wenxi He, Wei Sun
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引用次数: 0

摘要

背景:APC抑制域2 (Suppressor APC domain containing 2, SAPCD2)参与细胞周期调控,其mRNA水平在肿瘤组织中较高。但是,SAPCD2在癌症发展中的功能尚不清楚。目的:制备SAPCD2特异性小鼠单克隆抗体(mab),阐明SAPCD2在胃癌发生发展中的作用。方法:收集纯化的SAPCD2-GST免疫BALB/c小鼠脾细胞,与骨髓瘤细胞融合,获得单克隆抗体杂交瘤。制备了一组具有高特异性和敏感性的SAPCD2单克隆抗体,并通过IHC、WB、IP、IF和ELISA对其进行了鉴定。通过免疫组织化学分析,对228例胃粘膜病变临床标本(包括癌前病变和胃癌标本)中SAPCD2的表达进行了评价。结果:我们在真核细胞中鉴定出一个高度特异和敏感的s12克隆,并进行了免疫组化分析。我们发现107例胃癌组织中有81.3%是SAPCD2阳性,而在匹配的邻近正常组织中这一比例为26.2% (p结论:总之,我们描述了一种新的针对SAPCD2的单克隆抗体,这种抗体在胃癌组织中高表达,可能作为胃癌发展的早期临床标志物的基础。
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The Generation and Application of Monoclonal Antibodies to Detect SAPCD2 Expression in Precancerous and Malignant Gastric Lesions.

Background: Suppressor APC domain containing 2 (SAPCD2) is involved in cell cycle regulation and its mRNA levels are higher in cancer tissues. But, the function of SAPCD2 in cancer development remains unclear.

Objective: To generate mouse monoclonal antibodies (mAbs) specific to SAPCD2 and thus clarify the function of SAPCD2 in the development of gastric carcinoma (GC). Methods: Purified SAPCD2-GST immunized BALB/c mouse spleen cells were collected and fused with myeloma cells to obtain monoclonal antibody hybridoma. A group of monoclonal antibodies exhibiting high specificity and sensitivity against SAPCD2 has been generated and characterized by IHC, WB, IP, IF, and ELISA. By immunohistochemical analysis, the SAPCD2 expression was evaluated in 228 clinical samples of gastric mucosal lesions, including precancerous lesions and GC samples.

Results: We identified a highly specific and sensitive clone of s12 in eukaryotic cells and performed an IHC analysis. We found that 81.3% of 107 GC tissues were SAPCD2-positive, compared with the 26.2% in the matched adjacent normal-appearing tissues (P<0.001). Furthermore, among the 121 gastritis tissues, SAPCD2 was overexpressed in precancerous gastric lesions such as dysplasia (Dys, 78.9%), intestinal metaplasia (IM, 44.7%), and chronic atrophic gastritis (CAG, 6.1%) compared with that in chronic non-atrophic gastritis (CNAG, 3.2%) (P<0.001). The SAPCD2-positivity rate was 81.3% in GC, suggesting that the expression of SAPCD2 increased with the severity of the lesion (P<0.001).

Conclusion: In summary, we have described novel monoclonal antibodies against SAPCD2, which are highly expressed in GC tissues and may serve as the basis for an early clinical marker for GC development.

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来源期刊
Iranian Journal of Immunology
Iranian Journal of Immunology Medicine-Immunology and Allergy
CiteScore
1.60
自引率
0.00%
发文量
50
审稿时长
12 weeks
期刊介绍: The Iranian Journal of Immunology (I.J.I) is an internationally disseminated peer-reviewed publication and publishes a broad range of experimental and theoretical studies concerned with all aspects of immunology.
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